青海、陕西部分地区禾谷孢囊线虫rDNA-ITS-RFLP的特征分析

rDNA-ITS restriction fragment length polymorphism of cereal cyst nematodes in some regions of Qinghai,Shaanxi and Henan Province,China

采自我国青海、陕西等省地11个寄生小麦的孢囊线虫群体经形态学鉴定为禾谷孢囊线虫(Heterodera avenae)。用PCR技术扩增获得的rDNA-ITS片段长度约为1 060 bp。用HinfⅠ、TaqⅠ、HpaⅡ、HaeⅢ、PstⅠ、AluⅠ等6种限制性内切酶酶切ITS扩增产物;青海10群体YBT10A、HY65A、HY61B、ZHZ162B、HY5B、HHX8A、GH132A、HY92A、HY127B、DT142A的6个酶的RFLP图谱完全一致;陕西YL4A群体的HaeⅢ、HinfⅠ和HpaⅡ酶切结果与青海群体的上述3种酶切结果相同,但AluⅠ和PstⅠ的酶切结果比青海孢囊线虫群体都多1条1 060 bp片段,而YL4A群体TaqⅠ酶切结果较为复杂。对比已知Avenae组成员RFLP图谱,青海群体与北京房山H.avenae群体的RFLP图谱一致;而陕西YL4A的AluⅠ图谱与H.avenae法国群体一致,PstⅠ和TaqⅠ却与Avenae组成员已知酶切结果均不一致。河南3个禾谷孢囊线虫群体除TaqⅠ酶切结果比青海CCN群体多1条520 bp片段以外,其他5种酶的RFLP都一致,而与澳大利亚的禾谷孢囊线虫H.avenae群体的RFLP图谱一致。

Eleven populations of cereal cyst nematode（CCN） from wheat were collected in collected in Qinghai and Shaanxi Provinc,all morphologically identified as Heterodera avenae.The rDNA-ITS regions of the populations were amplified with the universal primers AB28 and TW81,a fragment of approximately 1 060 bp was yielded.The fragments were digested with Hinf Ⅰ,TaqⅠ,Hpa Ⅱ,Hae Ⅲ,Pst Ⅰ and Alu Ⅰ.The identical digestion patterns were obtained from CCN populations（YBT10A,HY65A,HY61B,ZHZ162B,HY5B,HHX8A,GH132A,HY92A,HY127B and DT142A） of Qinghai and which were the same species.The digestion patterns were the same between YL4A population of Shaanxi and the ones of Qinghai by Hae Ⅲ,Hinf Ⅰ and Hpa Ⅱ.The digestion patterns of Pst Ⅰ and Alu Ⅰ showed that the YL4A CCN population was differed from the ones of Qinghai,because it had an extra 1 060 bp fragment.The Taq Ⅰ restriction pattern of YL4A CCN population was complex.The obvious difference could be observed in the Taq Ⅰ RFLP pattern,a 520 bp fragment was not presented in the Qinghai CCN populations but in the Henan populations.According to the known conclusions,the analysis results restriction profiles showed that 10 populations of Qinghai were the identical RFLP patterns of H.avenae from China.Alu Ⅰ profile of YL4A population was the same as H.avenae from France,then digestion of YL4A population by Pst Ⅰ and Taq Ⅰ were different with either members of Avenae group because of its complex profiles.The Henan CCN populations were the similar as Qinghai ones except RFLP profile by Taq Ⅰ,and identical with H.avenae from Australia.